Decrease in the Time-Dependent Difference in Urinary Excretion of Furosemide with Age

The influence of age on the time-dependent difference in urinary excretion of furosemide, a loop diuretic agent, was examined in this longitudinal study. Male Wistar rats were maintained under conditions of light from 07:00 to 19:00 h and dark from 19:00 to 07:00 h. Furosemide (30 mg/kg) was given orally at 12:00 h (day trial) or 00:00 h (night trial) to rats at 3 months of age, and urine was collected for 8 h after dosage. Thereafter, the identical protocol was repeated using the same animals at 6, 9, 12, 15, 18, and 21 months of age. The urinary excretion of furosemide was significantly greater in the day than in the night trial at 3 months of age. Such a time-dependent difference was observed for up to 15 months, but disappeared at 18 and 21 months of age. The time-dependent difference in urinary excretion of furosemide (day trial — night trial) decreased gradually throughout the observation period of the study. These results suggest that the time-dependent difference in the urinary excretion of furosemide diminishes during the aging process and disappears by 18 months of age in male Wistar rats.     

Altered cardiovascular responses to chronic angiotensin II infusion in aged rats

In this work we determined by telemetry the cardiovascular effects produced by Ang II infusion on blood pressure (BP) and heart rate (HR) in aged rats. Male Wistar aged (48-52 weeks) and young (12 weeks) rats were used. Ang II (6 microg/h, young, n=6; aged, n=6) or vehicle (0.9% NaCl 1 microl/h, young, n=4; aged, n=5) were infused subcutaneously for 7 days, using osmotic mini-pump. The basal diurnal and nocturnal BP values were higher in aged rats (day: 98+/-0.3 mm Hg, night: 104+/-0.4 mm Hg) than in the young rats (day: 92+/-0.2 mm Hg, night: 99+/-0.2 mm Hg). In contrast, the basal diurnal and nocturnal HR values were significantly smaller in the aged rats. Ang II infusion produced a greater increase in the diurnal BP in the aged rats (Delta MAP=37+/-1.8 mm Hg) compared to the young ones (Delta MAP=30+/-3.5 mm Hg). In contrast, the nocturnal MAP increase was similar in both groups (young rats; Delta MAP=22+/-3.0 mm Hg, aged rats; Delta MAP=24+/-2.6 mm Hg). During Ang II infusion HR decreased transiently in the young rats. An opposite trend was observed in the aged rats. Ang II infusion also inverted the BP circadian rhythm, in both groups. No changes in HR circadian rhythm were observed. These differences suggest that the aging process alters in a different way Ang II-sensitive neural pathways involved in the control of autonomic activity.     

Development of progesterone dependency in the appearance of the nocturnal prolactin surge in immature rats

Basal concentrations of plasma prolactin in immature, Wistar-Imamichi strain rats at 25, 28 and 31 days of age were 5-12 ng/ml and no prolactin surges were observed in intact immature rats. Plasma progesterone values ranged from 5 to 9 ng/ml, while plasma oestradiol concentrations increased from 11 to 27 pg/ml between 25 and 31 days of age. When oestradiol was administered to ovariectomized 25- or 28-day-old rats by s.c. insertion of an implant, plasma prolactin concentrations at 05:00 and 12:00 h were similarly elevated 3 days after the operation. Oestradiol did not induce a nocturnal prolactin surge. The progesterone implants in ovariectomized rats at 28 days of age or on the first day of oestrus increased plasma prolactin values at 05:00 h. The magnitude of the progesterone-induced prolactin surge was greater when progesterone was given closer to the time of the first ovulation (about 34 days old). Pretreatment with oestradiol amplified the progesterone-induced prolactin surge. Mechanisms causing nocturnal prolactin surges are more sensitive to, and respond over a longer time period, to progesterone in pubertal rats than in adult animals. The results suggest that progesterone initiates the nocturnal surge of prolactin release and that oestradiol can amplify the effects of progesterone.     

Effects of male rat urine on norepinephrine levels in the accessory olfactory bulb of young and aged female rats.

Norepinephrine (NE) concentrations in the accessory olfactory bulbs (AOB) of young (4-5 months) and old (25-26 months) ovariectomized Sprague-Dawley rats, which were implanted with a 17 beta-estradiol silastic capsule and then exposed to male rat urine, were studied. The unilateral vomeronasal organ was removed in all rats one week before exposure to the urine stimulation. The NE level in the AOB of this surgical side served as the control. Urine collected from young adult male rats was poured into the female's cage at 12:00h and the animals were sacrificed before and 1, 2, or 3 hours after the male urine was given. The NE basal levels in the AOB of young rats decreased significantly at 14:00h compared to those at 12:00h, while no obvious changes in the NE concentrations were observed in the AOB of old rats during the same period. Two hours after continuous exposure to male urine, the NE concentrations in the AOB of young rats markedly increased, but no similar response occurred in the old rats. These data suggest that the noradrenergic functions in the AOB under basal conditions as well as in response to pheromonal stimulation may be modified with increasing age.     

Sleep and sleepiness among working and non-working high school evening students

The aim of this study was to evaluate patterns of sleepiness, comparing working and non-working students. The study was conducted on high school students attending evening classes (19:00-22:30 h) at a public school in S?o Paulo, Brazil. The study group consisted of working (n=51) and non-working (n=41) students, aged 14-21 yrs. The students answered a questionnaire about working and living conditions and reported health symptoms and diseases. For seven consecutive days, actigraphy measurements were recorded, and the students also filled in a sleep diary. Sleepiness ratings were given six times per day, including upon waking and at bedtime, using the Karolinska Sleepiness Scale. Statistical analyses included three-way ANOVA and t-test. The mean sleep duration during weekdays was shorter among workers (7.2 h) than non-workers (8.8 h) (t=4.34; p<.01). The mean duration of night awakenings was longer among workers on Tuesdays and Wednesdays (28.2 min) and shorter on Mondays (24.2 min) (t=2.57; p=.03). Among workers, mean napping duration was longer on Mondays and Tuesdays (89.9 min) (t=2.27; p=.03) but shorter on Fridays and Sundays (31.4 min) (t=3.13; p=.03). Sleep efficiency was lower on Fridays among non-workers. Working students were moderately sleepier than non-workers during the week and also during class on specific days: Mondays (13:00-15:00 h), Wednesdays (19:00-22:00 h), and Fridays (22:00-00:59 h). The study found that daytime sleepiness of workers is moderately higher in the evening. This might be due to a work effect, reducing the available time for sleep and shortening the sleep duration. Sleepiness and shorter sleep duration can have a negative impact on the quality of life and school development of high school students.     

Integrative pattern of vasomotor efficiency in SHR during ontogenesis

Numerous studies concerning the cardiovascular system in SHR often yield controversial data. The background of this diversity has various roots, ranging from different vascular segments or areas studied up to the different age of experimental animals. Our study aimed to follow the BP as an integrated response of vascular system. This approach was justified since stabilized cardiac output in SHR was proved till 1 year of age. The groups of male SHR (aged 3, 5, 9, 17 and 52 weeks) and age-matched Wistar rats were used. Significant basal BP difference between SHR and Wistar rats was found at 9 weeks of age and continued till the age of 52 weeks, reaching 189.6+/-11.9 mm Hg in SHR and 117.3+/-6.9 mm Hg in Wistar rats P<0.01 . The significant difference in BP increase to two doses of noradrenaline 0.1 microg and 1 microg between SHR and control rats was also found at the age of 9 weeks. At 52 weeks the BP increment to two doses of noradrenaline was in SHR 19.7+/-2.0 mm Hg and 60.5+/-3.9 mm Hg and in Wistar rats 7.4+/-1.9 mm Hg and 40.5+/-3.2 mm Hg P<0.01 . The hypotensive response to acetylcholine 0.1 microg, 1 microg and 10 microg in SHR was enhanced at 17 weeks of age only and this amplification persisted till the age of 52 weeks. In 52-week-old SHR the hypotensive response to three doses was 69.9+/-10.2 mm Hg, 87.5+/-11.8 mm Hg and 103.4+/-10.6 mm Hg, while in Wistar rats it was 37.4+/-4.2 mm Hg P<0.01 , 62.3+/-3.5 mm Hg P<0.01 and 73.5+/-2.8 mm Hg P<0.05 . In conclusion, the efficiency of cardiovascular system of SHR to respond to noradrenaline was already enhanced from 9 weeks of age, whereas the response to acetylcholine was not augmented before the age of 17 weeks.     

Does long-term experimental antiorthostasis lead to cardiovascular deconditioning in the rat?

Microgravity or simulated microgravity induces acute and chronic cardiovascular responses, whose mechanism is pivotal for understanding of physiological adaptation and pathophysiological consequences. We investigated hemodynamic responses of conscious Wistar rats to 45? head-down tilt (HDT) for 7 days. Arterial blood pressure (BP) was recorded by telemetry. Heart rate (HR), spectral properties and the spontaneous baroreflex sensitivity (sBRS) were calculated. Head-up tilt (HUT) was applied for 2 h before and after HDT to assess the degree of any possible cardiovascular deconditioning. Horizontal control BP and HR were 112.5+/-2.8 mmHg and 344.7+/-10 bpm, respectively. HDT elicited an elevation in BP and HR by 8.3 % and 8.8 %, respectively, in less than 1 h. These elevations in BP and HR were maintained for 2 and 3 days, respectively, and then normalized. Heart rate variability was unchanged, while sBRS was permanently reduced from the beginning of HDT (1.01+/-0.08 vs. 0.74+/-0.05 ms/mmHg). HUT tests before and after HDT resulted in BP elevations (6.9 vs. 11.6 %) and sBRS reduction (0.44 vs. 0.37 ms/mmHg), respectively. The pressor response during the post-HDT HUT test was accompanied by tachycardia (13.7 %). In conclusion, chronic HDT does not lead to symptoms of cardiovascular deconditioning. However the depressed sBRS and tachycardic response seen during the post-HDT HUT test may indicate disturbances in cardiovascular control.     

Fluctuations in peripheral serum testosterone levels within a day, with age and by sexual stimulation in male beagle dogs bred indoors

Fluctuations in serum levels of testosterone (T) within a day, both with age and as a result of sexual stimulation, were examined in male beagle dogs. Eighty male dogs aged between 3 months and 16 years and bred in our laboratory were used under strictly controlled breeding conditions (temperature: 22 +/- 1 degree C, relative humidity: 55 +/- 5%, lighting time: 8:00 a.m.-8:00 p.m.). The level of T was measured by an RIA method. In order to examine the fluctuation in T level within a day, blood samples were obtained at 0:00, 6:00, 12:00 and 18:00 in each of five dogs aged 1.7 and 2.1 years. T levels fluctuated with a regular pattern that was lowest at 12:00, and increased to a peak at 18:00-6:00, thereafter decreasing until 12:00. In order to examine the change in T level with age, blood samples were obtained at 9:00, 12:00 and 16:00 from 70 dogs aged between 3 months and 16 years. The regular diurnal pattern of T level was usually seen, and the levels at 12:00 were always low and did not fluctuate at any age except for 6 months, and 13, 14 and 16 years. The T level at 9:00 increased to reach a peak at 4 years, whereas that at 16:00 did so at 2 years. T levels at 9:00 were significantly higher at 4-12 and 14 years than at 3 months, and were higher at 4 years than at 9 months.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gastric Potential Difference in Fasted Rats: Circadian Studies

The pathophysiology of gastroduodenal ulcer disease remains the subject of intense research and controversy. One model of gastric ulcerogenesis implicates a disruption of complementary circadian rhythms between protective and destructive factors. Parallel circadian rhythms have been reported between acid secretion and gastric potential difference (PD) in in vitro models. The purpose of this study was to investigate the circadian measurements of PD, a parameter of intact gastric mucosal function and thus a putative parameter of gastric protection, in intact, fasted, anesthetized rats. Sixty-four male Sprague-Dawley rats were acclimatized in sound-attenuating, lightproof chambers for 3 weeks on a 12:12-h light-dark schedule. Eight rats were fasted 18 h before being sampled at each of eight times on the circadian clock (01:00, 04:00, 07:00, 10:00, 13:00, 16:00, 19:00. and 22:00 hours after lights on) (HALO). In each rat, after anesthesia (ketamine/ acepromazine) and laparotomy, the tip of a catheter (pre-filled with KC1 agar) was passed into the gastric corpus through the duodenum. The tip of a second KC1-agar catheter was placed within the peritoneal cavity. The position of the intragas-tric catheter was gently adjusted for obtaining the highest stable PD reading. The data showed significantly higher values at 07:00 and 10:00 HALO. The lowest value was at 13:00 HALO. The difference between high (10:00 HALO) and low (13:00 HALO) values was 4.5 mV or 13% of the mean. This difference was highly significant (p = 0.003) Analysis of variance showed that the values at 07:00 and 10:00 HALO were significantly higher than the values at 01:00, 13:00, and 16:00 HALO. Thus, the existence of a circadian rhythm in gastric PD is supported.     

Variations in the uptake of 3H-norepinephrine during the rat estrous cycle

The rates of ³H-norepinephrine (³H-NE) uptake into isolated nuclei-free homogenates from diencephalon were examined in freely cycling female rats of all phases of the estrous cycle. Four time points were investigated in each phase: 12:00, 14:00, 16:00 and 18:00 h. The proestrus and estrus animals displayed no change in uptake rates across the four time points. However, the metestrus and diestrus groups were found to undergo hour to hour variations in uptake rates. In the diestrus group, a significant rise in uptake of 22% was observed from 12:00 to 16:00 h. This increase was followed by a decrease of 21% by 18:00 h as compared to 16:00 h levels. Similarly, a slight increase in uptake was observed in the metustrus animals from 12:00 to 16:00 h. At 18:00 h a 40% decrease from the 16:00 h value was observed. An average of the mean levels of uptake of all phases was calculated for each time point. At 16:00 h an increase in the mean uptake of about 20% was observed when compared to either the 12:00 or the 18:00 h values. Since transmitter reuotake is the major means of terminating receptor activation or inhibition, it is suggested that the above variations in NE uptake may modulate synaptic activity and play a role in the regulation of behavioral and physiological aspects of the estrous cycle.     

Sleep length as a function of morning shift-start time in irregular shift schedules for train drivers: self-rated health and individual differences

Forty-six male train drivers (mean age = 46.5, SD = 5.1) were recruited to participate in a diary study for 14 consecutive days with questions about their sleep and working hours. A polynomial mixed-effect regression model showed a curvilinear relation ( p < .001) between shift-start time and sleep duration for shifts starting at 03:00-12:00 hand with a near linear increase for ones starting between 04:30 and 09:00 h of approximately 0.7 h for every 1 h the shift was delayed. The longest sleeps were estimated at approximately 8 h before shifts that started at approximately 10:00 h. The shortest sleeps were found for shifts that started before 04:30 h and were estimated at approximately 5 h. Individual differences were estimated with a random-effect standard deviation of 0.51 h, independent of shift start time ( p = .005). One-half of the between-subject variance was explained by subjective health. A one-step decrease in health was associated with a 26 min increase in sleep length. The results have practical implications for constructing shift schedules. Early morning shifts reduced sleep length substantially and should be mixed with later start hours to avoid the accumulation of sleep dept. Delaying the shift-start past 10:00 h had little effect on sleep opportunity; however, delaying shift-start to between 04:30 and 9:00 h had a strong impact on sleep length, with 70% of the extra time used for sleep, suggesting large positive effects for this range of shift-start times.     

Norepinephrine levels in the vomeronasal system and their responses to male urine in young and aged female rats

Norepinephrine (NE) levels in brain areas of the vomeronasal system in young (4-5 months) and aged (25-26 months) ovariectomized Sprague-Dawley rats, which were implanted with a 17 beta-estradiol silastic capsule and then exposed to male rat urine, were investigated. The unilateral vomeronasal organ was removed in all rats one week before exposure to urine stimulation. NE levels in the medial nucleus of the amygdala (MA), medial preoptic area (MPOA), ventromedial nucleus of hypothalamus (VMH) and bed nucleus of stria terminalis (BST) were measured. NE concentrations in these brain areas of the surgical side served as the control. Urine collected from young adult male rats was poured into the female's cage at 12:00h and the animals were sacrificed before and 1, 2, or 3 hours after the male urine was given. The NE basal levels in the MA and MPOA of young rats decreased significantly from 13:00h to 15:00h, and those in young rat VMH declined markedly from 13:00h to 14:00h compared to those at 12:00h. No marked alterations in NE basal levels in young rat BST were found. In contrast, no obvious changes in the NE concentrations were observed in these brain areas of old rats. Continuous exposure to male urine did not affect the NE levels in any of these brain areas of young and aged rats. We concluded that (1) the time-dependent fluctuation of the NE basal levels in some brain areas of the vomeronasal system in female rats is age-related, and (2) the NE in all these nuclei of the vomeronasal system is not involved in pheromone-induced effects.     

Circadian variations of serotonin in plasma and different brain regions of rats

Most of the physiological processes that take place in the organism follow a circadian rhythm. Serotonin is one of the most important neurotransmitters in our nervous system, and has been strongly implicated in the regulation on the mammalian circadian clock, located in the suprachiasmatic nuclei (SCN). The present study analysed the levels of serotonin over a period of 24 h in the plasma and in different brain regions. The model used was of male Wistar rats, 14 +/- 2 weeks of age (n = 120), maintained under conditions of 12 h light and 12 h dark, and food and water ad libitum. The serotonin levels were measured by ELISA every hour at night (20:00-08:00 h) and every 4 h during the daytime (08:00-20:00 h). Ours results show that the maximum levels of serotonin in plasma were obtained at 09:00 and 22:00 and a minor peak at 01:00 h. In hypothalamus there was a significant peak at 22:00 and two minor peaks at 17:00 and 02:00 h; the same occurred in hippocampus with a significant peak at 21:00, and two secondary peaks at 24:00 and 05:00 h; in cerebellum there were two peaks at 21:00 and 02:00 h, while in striatum and pineal there were peaks at 21:00 h and 23:00, respectively. In conclusion, the higher levels of serotonin were during the phase of darkness, which varies depending on the region in which it is measured.     

Differences in sleep, light, and circadian phase in offshore 18:00-06:00 h and 19:00-07:00 h shift workers

Complaints concerning sleep are high among those who work night shifts; this is in part due to the disturbed relationship between circadian phase and the timing of the sleep-wake cycle. Shift schedule, light exposure, and age are all known to affect adaptation to the night shift. This study investigated circadian phase, sleep, and light exposure in subjects working 18:00-06:00 h and 19:00-07:00 h schedules during summer (May-August). Ten men, aged 46+/-10 yrs (mean+/-SD), worked the 19:00-07:00 h shift schedule for two or three weeks offshore (58 degrees N). Seven men, mean age 41+/-12 yrs, worked the 18:00-06:00 h shift schedule for two weeks offshore (61 degrees N). Circadian phase was assessed by calculating the peak (acrophase) of the 6-sulphatoxymelatonin rhythm measured by radioimmunoassay of sequential urine samples collected for 72 h at the end of the night shift. Objective sleep and light exposure were assessed by actigraphy and subjective sleep diaries. Subjects working 18:00-06:00 h had a 6-sulphatoxymelatonin acrophase of 11.7+/-0.77 h (mean+/-SEM, decimal hours), whereas it was significantly later, 14.6+/-0.55 h (p=0.01), for adapted subjects working 19:00-07:00 h. Two subjects did not adapt to the 19:00-07:00 h night shift (6-sulphatoxymelatonin acrophases being 4.3+/-0.22 and 5.3+/-0.29 h). Actigraphy analysis of sleep duration showed significant differences (p=0.03), with a mean sleep duration for those working 19:00-07:00 h of 5.71+/-0.31 h compared to those working 18:00-06:00 h whose mean sleep duration was 6.64+/-0.33 h. There was a trend to higher morning light exposure (p=0.07) in the 19:00-07:00 h group. Circadian phase was later (delayed on average by 3 h) and objective sleep was shorter with the 19:00-07:00 h than the 18:00-06:00 h shift schedule. In these offshore conditions in summer, the earlier shift start and end time appears to favor daytime sleep.     

Effects of Amlodipine on Orcadian Rhythms in Blood Pressure, Heart Rate, and Motility: A Telemetric Study in Rats

In male Wistar rats [light (L): 07:00-19:00 h, dark (D): 19:00-07:00 h], the effects of the calcium channel blocker amlodipine (1, 3, 10 mg/kg i.p.) on blood pressure, heart rate, and motor activity were studied by telemetric monitoring. Amlodipine was injected either at 07:00 h or at 19:00 h. Systolic and diastolic blood pressure were dose-dependently decreased with more pronounced effects in the dark span, ED₅₀ values in D were about seven times lower than in L. In contrast, the dose-dependent increase in heart rate was more pronounced in L than in D. No significant effects of amlodipine were found on motor activity. The study gives evidence for a circadian phase-dependency in the cardiovascular effects amlodipine in rats.     

Follicle deviation and diurnal variation in circulating hormone concentrations in mares

The temporal relationships between follicle deviation and systemic hormone concentrations were studied in mares. Blood samples were obtained at 01:00, 07:00, 13:00, and 19:00 h from nine mares throughout an interovulatory interval. Diurnal variation in progesterone occurred on Days 4-12 and in LH on Days 4 and 5; the lowest concentration for both hormones was at 13:00 h. Ultrasonically observed deviation in the ovulatory follicular wave began on Day 15.7+/-0.5 (ovulation=Day 0). An increase (P<0.002) in LH began on Day 14 before the beginning of deviation, and an increase (P<0.05) in estradiol began at the beginning of deviation. Testosterone concentrations began to increase (P<0.05) 2 days after the beginning of deviation and reached maximum 1 day before the next ovulation. The beginning of deviation was encompassed by a decline (P<0.003) in cortisol concentrations, and the concentrations remained low during the preovulatory period.     

The features of sleep homeostasis in pregnant rats

To investigate the effects of short-term sleep deprivation on the sleep pattern during pregnancy, cortical and hippocampal EEG and locomotor activity were recorded within 24-hours in a "disk-over-water" paradigm in 18 Wistar rats. Rats were adapted to experimental situation and were able to move across the rotating disk without falling in water. Then a polysomnogram was recorded for 3 sequential days in the control group 1 (n = 12) without disk rotation. On the next day non-pregnant rats (experimental group 1, n = 6) were subjected to the sleep deprivation procedure with a pre-set program of disk rotation from 11:00 to 14:00 during 3 sequential days. Other 6 rats (experimental group 2) were subjected to sleep deprivation on the 5-7th day of pregnancy. EEG and locomotor activity were also constantly recorded during the sleep deprivation procedure. In control group 2 (n = 6, without sleep deprivation), a polysomnogram was recorded on the 5-7th day of pregnancy. As compared to non-pregnant rats, sleep intensity of pregnant rats increased during the first hours after the deprivation, and a considerable rebound of REM sleep took place. Sleep pattern during the off-light 12 hours remained unchanged. The results suggest that homeostatic compensation of sleep deprivation effects in rats on the first week of pregnancy is more efficient than in control non-pregnant animals.     

Effects of withholding food for 0-72 h on mating, pregnancy rate and pituitary function in female rats

Food was withheld from female rats for 0-72 h at various stages of the oestrous cycle. Withholding food for periods of 24 h ending at 12:00 h on the day of pro-oestrus reduced the mating rate from 61 to 25% (P less than 0.05) but not the pregnancy rate of those rats that mated. Fasting for 24 h ending at 18:00 h on the day of pro-oestrus reduced the pregnancy rate from 82 to 18% (P less than 0.05) without affecting the mating rate and a 48-h fast starting at 12:00 h on the day of pro-oestrus reduced the pregnancy rate from 82 to 25% (P less than 0.05). Withholding food for 23 h ending at 17:00 h on the day of pro-oestrus prevented the LH and prolactin surges normally present at 17:00 h on this day. The treatments had no apparent effect on the ability of the adenohypophysis to release LH in response to injections of GnRH. When ovariectomized female rats fasted for 0-72 h and given 2 injections of oestradiol dibenzoate to test the ability of the hypothalamus to respond to an increasing plasma oestradiol concentration by stimulating the release of LH, a fast for 24 h reduced and a fast for 72 h completely prevented LH release.     

Physiological pineal effects on female reproductive function of laboratory rats: prenatal development of pups, litter size and estrous cycle in middle age

The present study investigates whether and how the pineal or its hormone melatonin influences female reproductive functions, namely the litter size, prenatal development of offsprings, and estrous cyclicity, especially its age-related cessation in a non-seasonal breeder, the laboratory rat. Wistar rats were maintained under a 24 h light-dark (12Lratio12D) cycle. Female rats were divided into 3 groups: non-operated (NO), sham-operated (SX), and pinealectomized (PX). Surgeries were performed in 35-40 day-old females. Starting at an age between 70 days and 7 months, female rats of all 3 groups were repeatedly mated with intact males. PX mothers more frequently delivered pups with malformations (e.g., taillessness, hydronephrosis, 7 out of 1263 pups) than control rats (0/1323; p<0.007). In the first delivery at 3 months of age, but not at later ages, PX mothers delivered more pups of lower body weight than control animals (p<0.001). Examination of vaginal smears showed that almost all female rats of the NO, SX, and PX groups had 4-day estrous cyclicity when they were young-between 60 days and 5 months of age. At an age of 17 to 18 months, most female rats of the NO and SX groups showed irregular, continuously diestrous or pseudopregnancy-like patterns, and 4-day estrous cyclicity was found in only 10% of the NO or SX animals. In contrast, about 50% of the PX rats showed 4-day estrous cyclicity at this older age (p< 0.001). Melatonin, when added to drinking water (0.4 mg/L) for 16 days during the dark phase increased the frequency of diestrous phase, except in continuously diestrous rats and very few others. This melatonin effect was strong in PX rats but relatively weak in SX rats. In conclusion, the pineal hormone appears to influence various reproductive functions and developmental processes, especially pregnancy and the timing of reproductive aging in rats. The effects of pinealectomy are more prominent at an age of 60 to 80 days (i.e., shortly after puberty) and at the beginning of the cessation of cycles in middle-aged females.     

Relative bioavailability of copper in a copper-lysine complex or copper sulfate for ruminants as affected by feeding regimen

An experiment was conducted with 40 crossbred wether lambs to compare the relative bioavailability of Cu from a Cu-lysine complex with that from Cu sulfate under two feeding regimens. Liver biopsy samples were collected by a laparotomy technique and lambs were given 12 days to recover and adapt to individual pens. Lambs were fed either a control basal diet (9.8 mg kg⁻¹ Cu dry matter (DM) basis) or basal supplemented with 180 mg kg⁻¹ Cu as either reagent grade CuSO₄ · 5H₂O or a feed-grade Cu-lysine complex. There were 12 lambs fed control diet and 14 lambs fed each Cu-supplemented diet. Half of the lambs given each diet were fed 1.0 kg of their respective diet once daily at 08:00 h. The remaining half were fed 1.0 kg in equal portions (166 g) six times during a 24 h period at 04:00 h, 08:00 h, 12:00 h, 16:00 h, 20:00 h and 24:00 h. Lambs were fed experimental diets for 10 days, then killed and livers removed for Cu analysis. Lambs fed once daily accumulated more (P < 0.05) Cu in liver than those fed every 4 h. There was no Cu source × feeding regimen interaction (P > 0.10). Based on multiple linear regression slope ratio of log₁₀ transformed final liver Cu concentration on total intake of Cu (mg day⁻¹) with the initial liver biopsy Cu concentration as a covariate, bioavailability of Cu from Cu-lysine relative to Cu sulfate was 93.4% (P > 0.05) for combined feeding regimens.